Plant fermentation skin care composition and preparation method thereof

Abstract

The invention belongs to the technical field of skin care products, and relates to a plant fermentation skin care composition and a preparation method thereof. The composition is prepared from the following raw materials in percentage by weight: 0.5 to 1.5 percent of silybum marianum extract, 70 to 80 percent of rosemary fermentation essence and 20 to 30 percent of turmeric fermentation essence. The skin care composition inhibits skin aging through the following ways: 1, inhibiting KLK5 target expression, repairing skin barrier, inhibiting various skin inflammations, and preventing skin cell aging; 2, TIMP2 target expression is enhanced, and IV type collagen synthesis is promoted. And 3, the expression of an MMP3 target spot is inhibited, so that the degradation of various collagens such as III, IV and V types is reduced. And 4. Resisting oxidation, inhibiting activity of elastase, collagenase and hyaluronidase, inhibiting degradation of elastase, collagenase and hyaluronidase, and relieving skin aging.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care products, and more particularly, relates to a plant fermentation skin care composition and a preparation method thereof. Background Art

[0002] The skin is the outermost organ of the human body, consisting of the epidermis and the dermis. Among them, the epidermis directly exposed to the external environment is mainly composed of keratinocytes, with a thickness of about 0.05 - 1.5 millimeters. Under normal circumstances, keratinocytes continuously divide and differentiate, move towards the granular layer and the stratum corneum, produce intracellular lipids, and form a protective film.

[0003] Keratinocytes have a physical barrier function, which can reduce mechanical damage and the invasion of external substances. On the contrary, when keratinocytes continuously shed from the surface of the stratum corneum, the skin barrier is damaged, and various skin inflammations such as atopic dermatitis are likely to occur, reducing the skin repair ability and accelerating cell aging. Inhibiting the expression of KLK5 tissue kallikrein can effectively enhance the skin barrier and repair of the body, effectively inhibit the occurrence of various skin inflammations such as atopic dermatitis, and also has the effects of regulating skin moisture metabolism and soothing the skin.

[0004] Cell aging can occur not only in the epidermis but also in the dermis. Collagen and elastin in the dermis will accelerate the degradation and loss of collagen and elastin due to factors such as age growth, environmental stress, and ultraviolet radiation (such as free radicals), resulting in the breakage of the collagen peptide bonds and elastic nets that support the skin, the destruction of its helical network structure, the oxidation, atrophy, and collapse of the skin tissue, and the appearance of aging phenomena such as dryness, wrinkles, and loss of elasticity in the skin.

[0005] With the maturity and aging of the human skin, the content of hyaluronic acid gradually decreases, the skin moisturizing effect becomes poor, the nutritional metabolism becomes poor, the skin begins to become dry and water - deficient, loses luster and elasticity, and shows the phenomenon of relaxation and aging.

[0006] At present, the functions of anti - aging products on the market are not comprehensive. Therefore, there is great potential in developing a multi - dimensional anti - aging product that works from the inside out and transitions from the skin stratum corneum to the dermis.

[0007] Many plant extracts have various effects such as anti - inflammation, anti - allergy, and antioxidant. However, due to limitations in extraction processes and preparation methods, the active ingredients obtained from plant extracts prepared by traditional processes often fail to achieve the expected effects. In recent years, fermentation technology has shown good application prospects in the development of plant - based functional raw materials for cosmetics. Using fermentation technology to extract more abundant and smaller - molecular - weight functional active substances from green natural plant resources, enhancing the efficacy of plant extracts and reducing their toxic and side effects, not only helps more with the safety and effectiveness of plant extracts, but also greatly improves their functional effects.

[0008] Lactobacillus is a recognized probiotic in the human body, with characteristics such as high safety, fast growth rate, relatively clear metabolic mechanism, and few by-products. It is commonly used in the manufacture of yogurt, cheese, beer, pickles and other fermented foods, and has broad application prospects in the fields of bioengineering, industry and agriculture, and food processing. Summary of the Invention

[0009] The purpose of the present invention is to overcome the problems existing in the prior art and provide a plant-fermented skin care composition and its preparation method. The skin care composition of the present invention has the following advantages: First, it inhibits the expression of the KLK5 target, regulates the shedding of keratinocytes and the renewal of the epidermis, repairs the skin barrier, inhibits the occurrence of various skin inflammations, and prevents skin cell aging. Second, it enhances the expression of the TIMP2 target, promotes the synthesis of type IV collagen, regulates skin moisture, and promotes skin repair and regeneration. Third, it inhibits the expression of the MMP3 target, thereby reducing the degradation of various collagens such as type III, IV, and V, as well as proteoglycans and fibronectin. Fourth, it has antioxidant properties, inhibits the activities of elastase, collagenase, and hyaluronidase, thereby inhibiting the degradation of elastin, hyaluronic acid, and collagen, alleviating skin aging, and making the skin return to a youthful state.

[0010] The purpose of the present invention and the solution to its technical problems are achieved by adopting the following technical solutions.

[0011] One aspect of the present invention provides a plant-fermented skin care composition. Calculated by weight percentage, the composition includes the following raw materials in the following contents: 0.5-1.5% of silymarin extract, 70-80% of rosemary fermented essence, and 20-30% of turmeric fermented essence.

[0012] As a preferred embodiment of the present invention, the rosemary fermented essence is prepared by the following method:

[0013] S1: Take a certain amount of rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70-75%, and then stir at a temperature of 70-80°C for 2.5 h;

[0014] S2: After stirring, let it stand for 10 min and then filter by suction. Add 1,3-butanediol with a concentration of 70-75% again to the obtained suction filtrate.

[0015] As a preferred embodiment of the present invention, the rosemary fermentation extract in step S1 is prepared by the following method:

[0016] (1) After rinsing and sorting rosemary leaves with sterile water, blanch them in boiling water for 2 min, take them out and dry them, crush them, and pass through an 80-mesh sieve to obtain rosemary leaf powder for standby;

[0017] (2) Preparation of the culture medium: Take 20 - 100 g of rosemary leaf powder, add 1 L of distilled water, seal it to obtain the culture medium, and pasteurize the above - mentioned culture medium at 80 °C for 30 min;

[0018] (3) Inoculation and fermentation: Inoculate 1 - 10% of the fermentation strain Lactobacillus into the above - mentioned culture medium and ferment at 37 °C for 120 - 240 h;

[0019] (4) After the fermentation is completed, filter off the insoluble substances with filter paper to obtain the rosemary fermentation filtrate;

[0020] (5) Concentrate the obtained fermentation filtrate to an extract state with a rotary evaporator at a concentration temperature of 60 °C to obtain a rosemary fermentation extract with a water content of 5 - 10%.

[0021] As a preferred embodiment of the present invention, the addition amount of 1,3 - butanediol in step S1 is 7 times the total mass of the rosemary fermentation extract.

[0022] As a preferred embodiment of the present invention, the addition amount of 1,3 - butanediol in step S2 is 20 - 40% of the total mass of the filtrate after suction filtration.

[0023] As a preferred embodiment of the present invention, the turmeric fermentation essence is prepared according to the following method:

[0024] Take the turmeric fermentation extract, then add 1,3 - butanediol with a concentration of 70 - 80%, stir at 70 - 80 °C for 2.5 h, let it stand for 10 minutes after stirring, and finally perform suction filtration.

[0025] As a preferred embodiment of the present invention, the addition amount of 1,3 - butanediol is 7 times that of the turmeric fermentation extract.

[0026] As a preferred embodiment of the present invention, the turmeric fermentation extract is prepared according to the following method:

[0027] (1) After rinsing and arranging the turmeric root with sterile water, put it into boiling water for blanching for 2 min, take it out and dry the water, crush it, and pass it through an 80 - mesh sieve to obtain turmeric root powder for standby;

[0028] (2) Preparation of the culture medium: Take 20 g - 100 g of turmeric root powder, add 1 L of distilled water, seal it to obtain the culture medium, and pasteurize the above - mentioned culture medium at 80 °C for 30 min;

[0029] (3) Inoculation and fermentation: Inoculate 1 - 10% of the fermentation strain Lactobacillus into the above - mentioned culture medium and ferment at 37 °C for 120 - 240 h;

[0030] (4) After the fermentation is completed, filter off the insoluble substances with filter paper to obtain the turmeric fermentation filtrate;

[0031] (5) Concentrate the obtained fermentation filtrate to an extract state using a rotary evaporator at a concentration temperature of 60 °C to obtain a turmeric fermentation extract with a water content of 5-10%.

[0032] Another aspect of the present invention also provides a method for preparing the composition as described above, comprising the following steps: Weigh each component according to the formula amount, mix evenly, and centrifuge.

[0033] As a preferred embodiment of the present invention, the centrifugation conditions are: high-speed centrifugation, the centrifugation speed is 6000 rpm, the time is 10 min, and the temperature is 10 °C.

[0034] Another aspect of the present invention also provides an application of the composition as described above in the preparation of anti-aging skin care products.

[0035] By means of the above technical solutions, the present invention has at least the following advantages: 1. Through the different action mechanisms of different raw materials, the three raw material components in the composition of the present invention can effectively play a synergistic effect; 2. The composition of the present invention can inhibit the expression of the KLK5 target, regulate the shedding of keratinocytes and the renewal of the epidermis, repair the skin barrier, inhibit the occurrence of various skin inflammations, and prevent skin cell aging; 3. At the same time, it can also enhance the expression of the TIMP2 target, promote the synthesis of type IV collagen, regulate skin moisture, reduce skin dryness and roughness, promote skin repair and regeneration and antioxidant, inhibit the activities of elastase, collagenase, and hyaluronidase, relieve the effect of skin aging, and make the skin return to a young state.

[0036] The above description is only an overview of the technical solution of the present invention. In order to be able to understand the technical means of the present invention more clearly and implement it according to the content of the specification, the following will describe in detail with the preferred embodiments of the present invention as follows.

[0037] The preparation methods of the rosemary fermentation essence and turmeric fermentation essence described in the present invention are both obtained by using lactic acid bacteria fermentation technology. By using the enzymes metabolized by microorganisms in the fermentation engineering, the macromolecular active substances in rosemary and turmeric can be converted into small molecules that are more easily absorbed by the human body, further improving the active ingredients of rosemary and turmeric and enhancing the health care effects of rosemary and turmeric.

[0038] The lactic acid bacteria described in the present invention are Lactobacillus plantarum CGMCC NO.23158 or CGMCC NO.23159, which are preserved in the China General Microbiological Culture Collection Center. The address of the preservation unit is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen West Road, Chaoyang District, Beijing. The taxonomic name is Lactobacillus plantarum, and the preservation date is August 20, 2021. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Detection chart of the skin care effects of different compositions. Detailed implementation manners

[0040] In order to make the technical means, creative features, achieved purposes and functions of the present invention easy to understand, the following will combine the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention.

[0041] Unless otherwise specified, the 1,3-butanediol used in the present invention is of cosmetic grade and is provided by Guangzhou Ruoyuan Chemical Technology Co., Ltd.; Silybum marianum extract: solid state, with the content of silybin in Silybum marianum being (25-35%), provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.; Rosemary leaves and turmeric rhizomes are both provided by Anhui Shunhetang Chinese Herbal Pieces Co., Ltd.

[0042] Unless otherwise specified, the preparation method of rosemary fermentation extract produced by Lactobacillus fermentation in the embodiments of the present invention is as follows:

[0043] (1) Pretreatment of rosemary leaves: Rinse the sorted rosemary leaves with sterile water, put them into boiling water for blanching for 2 min after rinsing, take them out and dry the water, crush the rosemary leaves, and pass through an 80-mesh sieve to obtain rosemary leaf powder for standby.

[0044] (2) Preparation of rosemary culture medium: Take 60 g of rosemary leaves each, add 1 L of distilled water, seal, and pasteurize the above culture medium at 80 °C for 30 min.

[0045] (3) Inoculation and fermentation: Inoculate 10% of the fermentation strain into the culture medium at 37 °C and ferment for 240 h.

[0046] (4) After waiting for the fermentation to end, filter with filter paper to remove rosemary and bacteria to obtain the filtrate of Lactobacillus rosemary fermentation product.

[0047] (5) Concentrate the obtained filtrate to an extract state with a rotary evaporator at a temperature of 60 °C to obtain a rosemary fermentation extract with a water content of 10%.

[0048] Unless otherwise specified, the preparation method of turmeric fermentation product extract produced by Lactobacillus fermentation in the embodiments of the present invention is as follows:

[0049] (1) Pretreatment of turmeric roots: Rinse the sorted turmeric roots with sterile water, put them into boiling water for blanching for 2 min after rinsing, take them out and dry the water, crush the turmeric roots, and pass through an 80-mesh sieve to obtain turmeric root powder for standby.

[0050] (2) Preparation of turmeric root culture medium: Take 60 g of turmeric root powder, add 1 L of distilled water, seal it, and pasteurize the above culture medium at 80 °C for 30 min.

[0051] (3) Inoculation and fermentation: Inoculate 10% of the fermentation strain into the culture medium at 37 °C and ferment for 240 h.

[0052] (4) After waiting for the fermentation to end, filter with filter paper to remove turmeric and bacteria to obtain the filtrate of the Lactobacillus rosmarinus fermentation product.

[0053] (5) Concentrate the filtrate to an extract state with a rotary evaporator at a temperature of 60 °C to obtain a turmeric fermentation extract with a water content of 10%.

[0054] Unless otherwise specified, the method for detecting the collagenase inhibition rate involved in the embodiments of the present invention is as follows:

[0055] I. Experimental principle

[0056] Collagenase can hydrolyze FALGPA and has absorption at 335 nm. By measuring the OD value, the inhibition rate of the substrate on collagenase is reflected.

[0057] II. Equipment and reagents

[0058] Equipment: Constant temperature incubator, microplate reader, 96-well plates and other experimental consumables.

[0059] Reagents: Type I collagenase, PBS buffer, substrates: FALGPA, etc.

[0060] III. Experimental procedure

[0061] Prepare a 0.05 mol / L glycine buffer containing 0.4 mol / L NaCl and 0.01 mol / L CaCl2, and use this buffer to prepare a 1 U / mL type I collagenase solution and a 1 mmol / L N-[3-(2-furyl)acryloyl]-leucine-glycine-proline-alanine (FALGPA). Freeze-dried powder of the mung bean extract is prepared into a sample test solution with a certain concentration with deionized water. Then, mix 50 μL of the sample, 40 μL of the type I collagenase solution and 60 μL of the glycine buffer, incubate at 37 °C for 20 min, add 50 μL of the FALGPA solution, shake well, react at 37 °C for 30 min, and measure the absorbance at 335 nm with a microplate reader.

[0062] Use 1.2 mg / mL epigallocatechin gallate as a positive control, and determine the activity of collagenase according to the microplate reader reading.

[0063] Unless otherwise specified, the hyaluronidase activity inhibition experiment involved in the embodiments of the present invention is as follows:

[0064] I. Experimental principle

[0065] Hyaluronidase is a participant in type I allergic reactions. Hyaluronidase has a strong correlation with inflammation and allergy, as reported in research. Drugs that release histamine from various mast cells can regulate hyaluronidase activity, and some anti-allergic drugs have strong inhibitory effects on hyaluronidase activity. Therefore, inhibiting hyaluronidase activity is used as an indicator for studying anti-allergic effects.

[0066] II. Reagents and instruments

[0067] 1. Hyaluronidase: Concentration 1250 U / mL, freshly prepared and used immediately, not overnight, using acetate buffer as the solvent.

[0068] 2. Sodium hyaluronate: 0.5 mg / mL, prepared once and used multiple times, using acetate buffer as the solvent.

[0069] 3. Buffer: Solution A (11.55 mL of 0.2 mol / L acetic acid, glacial acetic acid dissolved in 1 L of distilled water) 4.8 mL, Solution B (16.4 g of 0.2 mol / L sodium acetate, anhydrous sodium acetate or 27.2 g of sodium acetate trihydrate dissolved in 1 L of distilled water) 45.2 mL, mixed and diluted to 100 mL to prepare an acetate buffer with pH = 5.6.

[0070] 4. Acetone solution: Mix 50 mL of 1.0 mol / L sodium carbonate solution and 3.5 mL of acetone solution evenly (prepared immediately before use).

[0071] P-DAB color reagent: Dissolve 0.8 g of p-dimethylaminobenzaldehyde in a mixture of 15 mL of concentrated hydrochloric acid and 15 mL of absolute ethanol and mix evenly.

[0072] 5. Calcium chloride solution CaCl2: 0.25 mM.

[0073] 6. NaOH solution: 0.4 M.

[0074] III. Calculation

[0075] Hyaluronidase inhibition rate (%) = (1 - (A - B) / (C - D)) * 100%

[0076] Wherein:

[0077] A: OD value of the sample solution of (hyaluronidase + sample + sodium hyaluronate);

[0078] B: OD value of the sample blank of (acetate buffer + sample + acetate buffer);

[0079] C: OD value of the control solution of (hyaluronidase + deionized water + sodium hyaluronate);

[0080] D: OD value of the control blank of (acetic acid buffer + deionized water + acetic acid buffer).

[0081] Unless otherwise specified, the process of the QPCR experiment (KLK5 / TIMP2) involved in the embodiments of the present invention is as follows:

[0082] I. Experimental principle

[0083] Real-time fluorescence quantitative PCR (RTFQ PCR) is to label and track the PCR products through fluorescent dyes or fluorescently labeled specific probes, monitor the reaction process in real time online, and analyze the products and calculate the initial concentration of the template of the sample to be tested in combination with the corresponding software.

[0084] II. Equipment and materials

[0085] QPCR instrument, cell incubator, centrifuge, laminar flow hood.

[0086] Culture medium, trypsin, serum, RNA extraction kit, reverse transcription kit, fluorescent dye.

[0087] III. Detection method

[0088] 1. Nucleic acid extraction: Extract the target DNA or RNA.

[0089] 2. Reverse transcription (RNA sample): Transcribe RNA into the corresponding cDNA.

[0090] 3. Pretreatment: Purify the extracted DNA / cDNA to remove contaminants.

[0091] 4. Primer and probe design: Design specific primers and fluorescent probes for specific binding to the target sequence.

[0092] 5. Prepare the reaction system: Prepare primers, probes, templates, polymerases, buffers, etc. according to the instructions.

[0093] 6. Perform PCR cycle: Load the reaction system into a thermal cycler and perform PCR cycle. Detect DNA denaturation, primer binding, amplification, and fluorescence signals, etc.

[0094] 7. Data analysis: Analyze the Ct value according to the change of fluorescence signal and calculate the relative quantity of the target sequence.

[0095] Unless otherwise specified, the reference standard for the free radical scavenging rate (DPPH) involved in the embodiments of the present invention is: the group standard T / SHRH006-2018 of the Shanghai Daily Chemical Industry Association.

[0096] Unless otherwise specified, the process for measuring the inhibitory activity of elastase involved in the embodiments of the present invention is as follows:

[0097] 1. Experimental principle

[0098] Elastase can hydrolyze N-methoxysuccinyl-ALA-ALA-PRO-VAL-p-nitroaniline to form a yellow product.

[0099] 2. Equipment and reagents

[0100] Equipment: Constant temperature incubator, microplate reader, 96-well plate, and other experimental consumables.

[0101] Reagents: Elastase, PBS buffer, substrate N-methoxysuccinyl-ALA-ALA-PRO-VAL-p-nitroaniline, DMSO, etc.

[0102] 3. Experimental process

[0103] Accurately weigh each natural product and dissolve it with DMSO to prepare a 20 mmol / L solution for standby. Take 135 (3U) μL of the prepared enzyme solution and 15 μL of the sample and add them to a 1.5 mL centrifuge tube. After reacting at 37 °C for 5 min, add 150 μL of the substrate N-methoxysuccinyl-ALA-ALA-PRO-VAL-p-nitroaniline (100 μM). At the same time, set up a 100% enzyme activity control group, a blank substrate group (i.e., replace the substrate with an equal volume of 10% DMSO (prepared with PBS)), and a blank control group. Set two parallel wells. After incubating at 37 °C for 60 min, centrifuge (10000 r / min, 5 min). Take 200 μL of the supernatant from each tube and transfer it to a 96-well microplate, and read the absorbance value of each well at a wavelength of 405 nm. The total reaction volume is 300 μL, and the final concentration of the drug in the reaction system is 1 mmol / L.

[0104] 4. Result evaluation

[0105] Calculate the inhibition rate of the drug on elastase according to the following formula:

[0106] Inhibition rate (%) = 100 × [1 - (A drug (subtracting the background) - A blank (control) (subtracting the background)) / A 100% enzyme well].

[0107] Example 1:

[0108] The total amount of the plant fermentation skin care composition in this embodiment is 1012 g, and the formula is as follows: turmeric fermentation essence, 200 g; rosemary fermentation essence, 800 g; silybum marianum extract, 12 g.

[0109] The preparation method of the plant composition is as follows:

[0110] 1. Extraction

[0111] 1.1 Take the rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70% which is 7 times the weight of the rosemary extract, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring and then filter by suction. Add 1,3-butanediol accounting for 25% of its weight to the obtained filtrate to obtain 800 g of rosemary fermentation essence.

[0112] 1.2 Take the turmeric fermentation extract, then add 1,3-butanediol with a concentration of 70% which is 7 times its weight, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring, and filter by suction to obtain 200 g of turmeric fermentation essence.

[0113] 1.3 Silybum marianum extract: solid state, silybin content (25 - 35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.), just take 12 g.

[0114] 2. Mixing

[0115] Mix the aforementioned 200 g of turmeric fermentation essence, 800 g of rosemary fermentation essence, and 12 g of silybum marianum extract evenly.

[0116] 3. Centrifugation

[0117] Centrifuge the above mixed solution with a high-speed centrifuge to obtain 1000 g of plant fermentation skin care product; the centrifugation parameters are: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0118] Example 2:

[0119] The total amount of the plant fermentation skin care composition in this embodiment is 1000 g, and the formula is as follows: turmeric fermentation essence, 220 g; rosemary fermentation essence, 780 g; silybum marianum extract, 12 g.

[0120] 1. Extraction

[0121] 1.1 Take the rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70% which is 7 times its weight, stir at 75 °C for 2.5 h, let it stand for 10 minutes after stirring and then filter by suction. Add 1,3-butanediol accounting for 20% of its weight to the obtained filtrate to obtain 780 g of rosemary fermentation essence.

[0122] 1.2 Take the turmeric fermentation extract, then add 1,3-butanediol with a concentration of 75% which is 7 times its weight, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring, and filter by suction to obtain 220 g of turmeric essence.

[0123] 1.3 Silybum marianum extract: solid state, silybin content (25 - 35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.), take 12 g.

[0124] 2 Mixing

[0125] Mix the aforementioned 200 g of turmeric fermentation essence, 800 g of rosemary fermentation essence, and 12 g of silybum marianum extract evenly.

[0126] 3 Centrifugation

[0127] Centrifuge the above mixture with a high-speed centrifuge to obtain 1000 g of plant fermentation skin care product; centrifugation parameters: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0128] Example 3:

[0129] The total amount of the plant fermentation skin care composition in this example is 1000 g, and the formula is as follows: turmeric fermentation essence, 210 g; rosemary fermentation essence, 790 g; silybum marianum extract, 12 g.

[0130] 1 Extraction

[0131] 1.1 Take the rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70% which is 7 times the weight of the rosemary extract, stir at 75 °C for 2.5 h, let it stand for 10 minutes after stirring and filter by suction, and add 1,3-butanediol with 20% of its weight to the obtained filtrate to obtain 790 g of rosemary fermentation essence.

[0132] 1.2 Take the turmeric fermentation extract, then add 1,3-butanediol with a concentration of 75% which is 7 times its weight, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring, and filter by suction to obtain 210 g of turmeric fermentation essence.

[0133] 1.3 Silybum marianum extract: solid state, silybin content (25 - 35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.), take 12 g.

[0134] 2 Mixing

[0135] Mix the aforementioned 200 g of turmeric fermentation essence, 800 g of rosemary fermentation essence, and 12 g of silybum marianum extract evenly.

[0136] 3 Centrifugation

[0137] Centrifuge the above-mentioned mixture with a high-speed centrifuge to obtain 1000 g of plant fermented skin care products; centrifugation parameters: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0138] Comparative Examples 1-3

[0139] Experimental purpose: To detect the skin care effects of different compositions. Referring to Example 1, make single variable changes (Comparative Examples 1-3)

[0140] Comparative Example 1:

[0141] The total amount of the plant fermented skin care composition in this comparative example is 1000 g, and the formula is as follows: turmeric fermented essence, 200 g; rosemary fermented essence, 800 g;

[0142] 1 Extraction

[0143] 1.1 Take rosemary fermented extract, then add 1,3-butanediol with a concentration of 70% and a weight 7 times that of the extract, stir at 75 °C for 2.5 h, let it stand for 10 minutes after stirring and then filter by suction. Add 1,3-butanediol with a weight 20% of the filtrate to obtain 800 g of rosemary fermented essence.

[0144] 1.2 Take turmeric fermented extract, then add 1,3-butanediol with a concentration of 75% and a weight 7 times that of the turmeric rhizome, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring, and filter by suction to obtain 200 g of turmeric fermented essence.

[0145] 2 Mixing

[0146] Mix the above-mentioned 200 g of turmeric fermented essence and 800 g of rosemary fermented essence evenly.

[0147] 3 Centrifugation

[0148] Centrifuge the above-mentioned mixture with a high-speed centrifuge to obtain 1000 g of plant fermented skin care products; centrifugation parameters: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0149] Comparative Example 2:

[0150] The total amount of the plant fermented skin care composition in this comparative example is 212 g, and the formula is as follows: turmeric fermented essence, 200 g; silybum marianum extract (total content 1.0%-1.5%), 12 g;

[0151] 1 Extraction

[0152] 1.1 Silybum marianum extract: solid state, silybin content in silybum marianum (25-35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.), take 12 g.

[0153] 1.2 Take the turmeric fermentation extract, then add 1,3-butanediol with a concentration of 75% and a weight 7 times that of the turmeric fermentation extract, stir at 70 °C for 2.5 h, let stand for 10 minutes after stirring, and filter to obtain 200 g of turmeric fermentation essence.

[0154] 2 Mix

[0155] Mix 200 g of the above-mentioned turmeric fermentation essence and 12 g of silymarin extract evenly.

[0156] 3 Centrifuge

[0157] Centrifuge the above mixture with a high-speed centrifuge to obtain 212 g of plant fermented skin care product; centrifugation parameters: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0158] Comparative Example 3:

[0159] The total amount of the plant fermented skin care composition in this comparative example is 812 g, and the formula is as follows: rosemary fermentation essence, 800 g; silymarin extract, 12 g;

[0160] 1 Extraction

[0161] 1.1 Silymarin extract: solid state, silybin content in silymarin (25 - 35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.), take 12 g.

[0162] 1.2 Take the rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70% and a weight 7 times that of the rosemary leaves, stir at 75 °C for 2.5 h, let stand for 10 minutes after stirring and filter, and add 1,3-butanediol with a weight 20% of the obtained filtrate to obtain 800 g of rosemary fermentation essence.

[0163] 2 Mix

[0164] Mix 200 g of the above-mentioned turmeric fermentation essence and 800 g of rosemary fermentation essence evenly.

[0165] 3 Centrifuge

[0166] Centrifuge the above mixture with a high-speed centrifuge to obtain 812 g of plant fermented skin care product; centrifugation parameters: rotation speed: 6000 rpm, time: 10 min, temperature: 10 °C.

[0167] Comparative Examples 4 - 6

[0168] Experimental purpose: Refer to Example 1, detect the efficacy of single components, and compare the efficacy of combinations 2 and 3 (Comparative Examples 1 - 3)

[0169] Comparative Example 4:

[0170] 1000 grams of silymarin extract in this combined formula.

[0171] 1 Extraction

[0172] 1.1 Silymarin extract: solid state, silybin content (25 - 35%) (provided by Shaanxi Jinkangtai Biotechnology Co., Ltd.). Take 12 g and dissolve it in 1000 grams of 77% butylene glycol, stir for 30 minutes, extract at 50 °C, and then perform suction filtration.

[0173] Comparative Example 5:

[0174] 800 grams of rosemary fermentation essence in this combined formula.

[0175] 1.1 Take rosemary fermentation extract, then add 7 times its weight of 1,3 - butylene glycol with a concentration of 70%, stir at 75 °C for 2.5 h, let it stand for 10 minutes after stirring and then perform suction filtration. Add 20% of its weight of 1,3 - butylene glycol to the obtained filtrate to make the butylene glycol concentration reach 77%, and suction filter to obtain 800 g of rosemary fermentation essence.

[0176] Comparative Example 6:

[0177] 200 grams of turmeric fermentation essence in this combined formula.

[0178] Take turmeric fermentation extract, then add 7 times its weight of 1,3 - butylene glycol with a concentration of 75%, stir at 70 °C for 2.5 h, let it stand for 10 minutes after stirring, and then suction filter to obtain 200 g of turmeric fermentation essence.

[0179] Experiment 1: Detection of Physicochemical Properties of Different Combinations

[0180] According to the above - mentioned method, the skin - care effects of the combinations in Examples 1 - 3 and Comparative Examples 1 - 6 were detected, and the results are shown in Table 1 and Figure 1 .

[0181] Table 1 Detection of Skin - care Effects of Different Combinations

[0182]

[0183]

[0184] Note: The detection concentration of KLK5 / TIMP2 / MMP3 is 0.2% (stock solution), which is the safe concentration, and the concentrations of other detection indicators are stock solutions.

[0185] From Table 1 and Figure 1 the results, it can be seen that compared with Comparative Examples 1 - 6, the combinations in Examples 1 - 3 of the present invention have higher elastase inhibition rates, collagenase inhibition rates, etc. Skin - care combinations lacking any one of these substances (Comparative Examples 1 - 6) cannot achieve the same effect.

[0186] Experiment 2: Detection of the clinical use effects of different compositions

[0187] To evaluate the clinical use effects of the compositions of the present invention, the following tests were conducted in this experiment. The specific operation method was as follows: 70 subjects were randomly selected, aged between 30 and 45, including 20 males and 50 females. The above subjects were randomly divided into 7 groups, with 10 subjects in each group. Before the test, photos were taken using a VISIA facial image analyzer to record the initial value A0 of the number of wrinkles. The compositions of Example 1 and Comparative Examples 1-6 were used for each group of subjects respectively. The usage time was after washing the face every morning and before going to bed. During the usage period, the subjects in each group maintained relatively consistent living and working habits. A follow-up visit was conducted on the 30th day, facial photos were taken using VISIA, and the number of wrinkles A1 was recorded. Subsequently, the improvement rate of facial wrinkles before and after use was statistically calculated, and the results are shown in Table 2.

[0188] Table 2 Wrinkles improvement rate

[0189]

[0190]

[0191] Thus, it can be seen that the plant anti-aging skin care composition of the present invention has significant and comprehensive effects. The plant fermentation skin care composition of the present invention has the following functions: First, it inhibits the expression of the KLK5 target, regulates the shedding of keratinocytes and the renewal of the epidermis, repairs the skin barrier, inhibits the occurrence of various skin inflammations, and prevents skin cell aging. Second, it enhances the expression of the TIMP2 target, promotes the synthesis of type IV collagen, regulates skin moisture, and promotes skin repair and regeneration. Third, it inhibits the expression of the MMP3 target, thereby reducing the degradation of various collagens such as type III, IV, and V, as well as proteoglycans and fibronectin. Fourth, it has antioxidant effects, inhibits the activities of elastase, collagenase, and hyaluronidase, thereby inhibiting the degradation of elastin, hyaluronic acid, and collagen, alleviating skin aging, and making the skin return to a youthful state. Moreover, the product uses the enzymes metabolized in the microbial fermentation technology process to convert the macromolecular active substances in rosemary and turmeric into small molecules that are more easily absorbed by the human body, further improving the active ingredients of rosemary and turmeric, enhancing the health care effects of rosemary and turmeric, and the product is safe and non-irritating.

[0192] The above are only the preferred embodiments of the present invention and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art, without departing from the scope of the technical solution of the present invention, can make some changes or modifications to the above-disclosed methods and technical contents to form equivalent embodiments of equivalent changes. However, as long as it does not depart from the content of the technical solution of the present invention, any simple modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A plant fermentation skin care composition, characterized in that, By weight percentage, the composition comprises raw materials in the following contents: 0.5-1.5% of silybum marianum extract, 70-80% of rosemary fermentation essence, and 20-30% of turmeric fermentation essence.

2. The composition according to claim 1, wherein The rosemary fermentation essence is prepared by the following method: S1: Take a certain amount of rosemary fermentation extract, then add 1,3-butanediol with a concentration of 70-75%, and then stir at a temperature of 70-80°C for 2.5 h; S2: After stirring, let it stand for 10 min and then filter by suction. Then add 1,3-butanediol with a concentration of 70-75% again to the obtained suction filtrate.

3. The composition according to claim 2, wherein The rosemary fermentation extract in step S1 is prepared by the following method: (1) After rinsing and tidying rosemary leaves with sterile water, blanch them in boiling water for 2 min, take them out and dry them in the air. Then crush them and pass through an 80-mesh sieve to obtain rosemary leaf powder for standby; (2) Preparation of the culture medium: Take 20-100 g of rosemary leaf powder, add 1 L of distilled water, seal it to obtain the culture medium, and pasteurize the above culture medium at 80°C for 30 min; (3) Inoculation and fermentation: Inoculate 1-10% of the fermentation strain Lactobacillus into the above culture medium and ferment at 37°C for 120-240 h; (4) After fermentation, filter off the insoluble matters with filter paper to obtain rosemary fermentation filtrate; (5) Concentrate the obtained fermentation filtrate to an extract state with a rotary evaporator at a concentration temperature of 60°C to obtain a rosemary fermentation extract with a water content of 5-10%.

4. The composition according to claim 2, wherein The addition amount of 1,3-butanediol in step S1 is 7 times the total mass of the rosemary fermentation extract.

5. The composition according to claim 2, wherein The addition amount of 1,3-butanediol in step S2 is 20-40% of the total mass of the suction filtrate.

6. The composition according to claim 1, wherein The turmeric fermentation essence is prepared by the following method: Take the turmeric fermentation extract, then add 1,3-butanediol with a concentration of 70-80%, stir at 70-80°C for 2.5 h, let it stand for 10 minutes after stirring, and finally filter by suction.

7. The composition according to claim 6, wherein The addition amount of 1,3-butanediol is 7 times that of the turmeric fermentation extract.

8. The composition according to claim 6, characterized in that, The turmeric fermentation extract is prepared by the following method: (1) After rinsing and tidying turmeric roots with sterile water, blanch them in boiling water for 2 min, take them out and dry them in the air. Then crush them and pass through an 80-mesh sieve to obtain turmeric root powder for standby; (2) Preparation of the culture medium: Take 20 g-100 g of turmeric root powder, add 1 L of distilled water, seal it to obtain the culture medium, and pasteurize the above culture medium at 80°C for 30 min; (3) Inoculation and fermentation: Inoculate 1-10% of the fermentation strain Lactobacillus into the above culture medium and ferment at 37°C for 120-240 h; (4) After fermentation, filter off the insoluble matters with filter paper to obtain turmeric fermentation filtrate; (5) Concentrate the obtained fermentation filtrate to an extract state with a rotary evaporator at a concentration temperature of 60°C to obtain a turmeric fermentation extract with a water content of 5-10%.

9. A method for preparing the composition according to any one of claims 1-8, characterized in that, It includes the following steps: Weigh each component according to the formula amount, mix them evenly, and then centrifuge.

10. The method according to claim 9, wherein The centrifugation conditions are: high-speed centrifugation, the centrifugation speed is 6000 rpm, the time is 10 min, and the temperature is 10°C.

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